Protocol for in vitro co-culture, proliferation, and cell cycle analyses of patient-derived leukemia cells

Jessica Parker; Sean Hockney; Carly Knill; David McDonald; Andrew Filby; Deepali Pal

STAR Protocols (Sep 2024)

Protocol for in vitro co-culture, proliferation, and cell cycle analyses of patient-derived leukemia cells

Jessica Parker,
Sean Hockney,
Carly Knill,
David McDonald,
Andrew Filby,
Deepali Pal

Affiliations

Jessica Parker: Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK
Sean Hockney: Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK
Carly Knill: Flow Cytometry Core Facility (FCCF), Newcastle University, Newcastle upon Tyne NE1 7RU, UK; Biosciences Institute, Innovation, Methodology and Application (IMA) Research Theme, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK
David McDonald: Flow Cytometry Core Facility (FCCF), Newcastle University, Newcastle upon Tyne NE1 7RU, UK; Biosciences Institute, Innovation, Methodology and Application (IMA) Research Theme, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK
Andrew Filby: Flow Cytometry Core Facility (FCCF), Newcastle University, Newcastle upon Tyne NE1 7RU, UK; Biosciences Institute, Innovation, Methodology and Application (IMA) Research Theme, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK
Deepali Pal: Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK; Wolfson Childhood Cancer Research Centre, Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Herschel Building Level 6, Brewery Lane, Newcastle upon Tyne NE1 7RU, UK; School of Cellular and Molecular Medicine, University of Bristol, Biomedical Sciences Building, University Walk, Bristol BS8 1TD, UK; Corresponding author

Journal volume & issue: Vol. 5, no. 3
p. 103202

Abstract

Read online

Summary: Leukemia niche impacts quiescence; however, culturing patient-derived samples ex vivo is technically challenging. Here, we present a protocol for in vitro co-culture of patient-derived xenograft acute lymphoblastic leukemia (PDX-ALL) cells with human mesenchymal stem cells (MSCs). We describe steps for labeling PDX-ALL cells with CellTrace Violet dye to demonstrate MSC-primed PDX-ALL cycling. We then detail procedures to identify MSC-primed G0/quiescent PDX-ALL cells via Hoechst-33342/Pyronin Y live cell cycle analysis.For complete details on the use and execution of this protocol, please refer to Pal et al.1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords